Composition for Lowering Blood Lipid and Elevating High-density Lipoprotein and Method for Manufacturing the Same

Abstract

The present invention discloses a composition for lowering blood lipid and elevating high-density lipoprotein and a method for manufacturing the same; the composition comprises monascin or ankaflavin, or a combination thereof; the manufacturing method comprises the steps of: treating a Monascus fermented product with acetone for three times; elevating the concentration of the Monascus fermented product by a process of decompress concentration; and extracting the monascin and the ankaflavin from the Monascus fermented product with a silica gel column chromatography, a Sephadex LH-20 column chromatography, the silica gel column chromatography, and a preparative high performance liquid chromatography sequentially.

1 . A composition for lowering blood lipid and elevating high-density lipoprotein, wherein the composition comprises monascin, which is a Monascus yellow pigment and extracted from a Monascus fermented product. 
     
     
         2 . The composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 1 , wherein an amount of the monascin taken by an adult is more than 2.4 mg per day. 
     
     
         3 . A method for manufacturing a composition for lowering blood lipid and elevating high-density lipoprotein, wherein the method comprises the steps of:
 (1) providing a Monascus fermented product;   (2) treating the Monascus fermented product with acetone for three times;   (3) elevating the concentration of the product obtained from the previous step by a process of decompress concentration in a specific temperature range;   (4) separating a pigment fraction from the product obtained from the previous step by a silica gel column chromatography;   (5) separating a yellow pigment fraction from the pigment fraction by a Sephadex LH-20 column chromatography;   (6) separating a fraction containing monascin and ankaflavin from the yellow pigment fraction by the silica gel column chromatography; and   (7) separating monascin from the fraction containing monascin and ankaflavin obtained from the previous step by a preparative high performance liquid chromatography (pre-HPLC).   
     
     
         4 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the ratio of the  Monascus  fermented product to the acetone in step (2) is 1:10˜1:50. 
     
     
         5 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the specific temperature range in step (3) is 40˜60. 
     
     
         6 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the step (4) further comprises the steps of:
 (4.1) adding the product obtained from step (3) to a silica gel column;   (4.2) adding a plurality of washing solutions to the silica gel column sequentially;   (4.3) collecting 12˜15 fractions flowing out from the silica gel column;   (4.4) analyzing the fractions collected in the previous step by a high performance liquid chromatography (HPLC) in combination with a photodiode-array (PDA), and collecting fractions containing monascin and ankaflavin; and   (4.5) mixing the fractions collected in the previous step to form the pigment fraction.   
     
     
         7 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 6 , wherein the plurality of washing solutions in step (4.2) are Hexane, Hexane:Ethanol=9:1, Hexane:Ethanol=8:2, Hexane:Ethanol=7:3, Hexane:Ethanol=1:1, and Ethanol sequentially. 
     
     
         8 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the step (5) further comprises the steps of:
 (5.1) adding the pigment fraction obtained from step (4) to a Sephadex LH-20 column;   (5.2) adding a washing solution to the Sephadex LH-20 column;   (5.3) collecting 3˜5 fractions flowing out from the Sephadex LH-20 column;   (5.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (5.5) mixing the fractions collected in the previous step to form the yellow pigment fraction.   
     
     
         9 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 8 , wherein the washing solution in step (5.2) is Methanol:Acetonitrile=9:1. 
     
     
         10 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the step (6) further comprises the steps of:
 (6.1) adding the yellow pigment fraction obtained from step (5) to a silica gel column;   (6.2) adding a plurality of washing solutions to the silica gel column sequentially;   (6.3) collecting 3˜5 fractions flowing out from the silica gel column;   (6.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (6.5) mixing the fractions collected in the previous step to form the fraction containing monascin and ankaflavin.   
     
     
         11 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 10 , wherein the plurality of washing solutions in step (6.2) are Dichloromethane:Ethanol=95:5, Dichloromethane:Ethanol=9:1, and Dichloromethane:Ethanol=4:1 sequentially. 
     
     
         12 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 3 , wherein the step (7) further comprises the steps of:
 (7.1) washing and separating the fraction containing monascin and ankaflavin obtained from step (6) by a C 18  column in combination with a HPLC and a washing solution;   (7.2) collecting 2 fractions flowing out from the C 18  column; and   (7.3) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin.   
     
     
         13 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 12 , wherein the washing solution in step (7.1) is Methanol:water=85:15. 
     
     
         14 . A composition for lowering blood lipid and elevating high-density lipoprotein, wherein the composition comprises ankaflavin, which is a Monascus yellow pigment and extracted from a Monascus fermented product. 
     
     
         15 . The composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 14 , wherein an amount of the ankaflavin taken by an adult is more than 0.6 mg per day. 
     
     
         16 . A method for manufacturing a composition for lowering blood lipid and elevating high-density lipoprotein, wherein the method comprises the steps of:
 (1) providing a  Monascus  fermented product;   (2) treating the  Monascus  fermented product with acetone for three times;   (3) elevating the concentration of the product obtained from the previous step by a process of decompress concentration in a specific temperature range;   (4) separating a pigment fraction from the product obtained from the previous step by a silica gel column chromatography;   (5) separating a yellow pigment fraction from the pigment fraction by a Sephadex LH-20 column chromatography;   (6) separating a fraction containing monascin and ankaflavin from the yellow pigment fraction by the silica gel column chromatography; and   (7) separating ankaflavin from the fraction containing monascin and ankaflavin obtained from the previous step by a pre-HPLC.   
     
     
         17 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the ratio of the  Monascus  fermented product to the acetone in step (2) is 1:10˜1:50. 
     
     
         18 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the specific temperature range in step (3) is 40˜60. 
     
     
         19 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the step (4) further comprises the steps of:
 (4.1) adding the product obtained from step (3) to a silica gel column;   (4.2) adding a plurality of washing solutions to the silica gel column sequentially;   (4.3) collecting 12˜15 fractions flowing out from the silica gel column;   (4.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (4.5) mixing the fractions collected in the previous step to form the pigment fraction.   
     
     
         20 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 19 , wherein the plurality of washing solutions in step (4.2) are Hexane, Hexane:Ethanol=9:1, Hexane:Ethanol=8:2, Hexane:Ethanol=7:3, Hexane:Ethanol=1:1, and Ethanol sequentially. 
     
     
         21 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the step (5) further comprises the steps of:
 (5.1) adding the pigment fraction obtained from step (4) to a Sephadex LH-20 column;   (5.2) adding a washing solution to the Sephadex LH-20 column;   (5.3) collecting 3˜5 fractions flowing out from the Sephadex LH-20 column;   (5.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (5.5) mixing the fractions collected in the previous step to form the yellow pigment fraction.   
     
     
         22 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 21 , wherein the washing solution in step (5.2) is Methanol:Acetonitrile=9:1. 
     
     
         23 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the step (6) further comprises the steps of:
 (6.1) adding the yellow pigment fraction obtained from step (5) to a silica gel column;   (6.2) adding a plurality of washing solutions to the silica gel column sequentially;   (6.3) collecting 3˜5 fractions flowing out from the silica gel column;   (6.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (6.5) mixing the fractions collected in the previous step to form the fraction containing monascin and ankaflavin.   
     
     
         24 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 23 , wherein the plurality of washing solutions in step (6.2) are Dichloromethane:Ethanol=95:5, Dichloromethane:Ethanol=9:1, and Dichloromethane:Ethanol=4:1 sequentially. 
     
     
         25 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 16 , wherein the step (7) further comprises the steps of:
 (7.1) washing and separating the fraction containing monascin and ankaflavin obtained from step (6) by a C 18  column in combination with a HPLC and a washing solution;   (7.2) collecting 2 fractions flowing out from the C 18  column; and   (7.3) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing ankaflavin.   
     
     
         26 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 25 , wherein the washing solution in step (7.1) is Methanol:water=85:15. 
     
     
         27 . A composition for lowering blood lipid and elevating high-density lipoprotein, wherein the composition comprises monascin and ankaflavin, which are both  Monascus  yellow pigments and extracted from a  Monascus  fermented product. 
     
     
         28 . The composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 27 , wherein the content ratio of the monascin to the ankaflavin is in a range from 1:1 to 10:1. 
     
     
         29 . The composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 27 , wherein the optimum content ratio of the monascin to the ankaflavin is 3.56:1. 
     
     
         30 . The composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 27 , wherein an amount of the monascin taken by an adult is more than 2.4 mg per day, and wherein an amount of the ankaflavin taken by the adult is more than 0.6 mg per day. 
     
     
         31 . A method for manufacturing a composition for lowering blood lipid and elevating high-density lipoprotein, wherein the method comprises the steps of:
 (1) providing a Monascus fermented product;   (2) treating the Monascus fermented product with acetone for three times;   (3) elevating the concentration of the product obtained from the previous step by a process of decompress concentration in a specific temperature range;   (4) separating a pigment fraction from the product obtained from the previous step by a silica gel column chromatography;   (5) separating a yellow pigment fraction from the pigment fraction by a Sephadex LH-20 column chromatography;   (6) separating a fraction containing monascin and ankaflavin from the yellow pigment fraction by the silica gel column chromatography;   (7) separating monascin and ankaflavin respectively from the fraction containing monascin and ankaflavin obtained from the previous step by a pre-HPLC; and   (8) mixing the monascin and the ankaflavin according to a specific ratio.   
     
     
         32 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the ratio of the Monascus fermented product to the acetone in step (2) is 1:10˜1:50. 
     
     
         33 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the specific temperature range in step (3) is 40˜60. 
     
     
         34 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the step (4) further comprises the steps of:
 (4.1) adding the product obtained from step (3) to a silica gel column;   (4.2) adding a plurality of washing solutions to the silica gel column sequentially;   (4.3) collecting 12˜15 fractions flowing out from the silica gel column;   (4.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (4.5) mixing the fractions collected in the previous step to form the pigment fraction.   
     
     
         35 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 34 , wherein the plurality of washing solutions in step (4.2) are Hexane, Hexane:Ethanol=9:1, Hexane:Ethanol=8:2, Hexane:Ethanol=7:3, Hexane:Ethanol=1:1, and Ethanol sequentially. 
     
     
         36 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the step (5) further comprises the steps of:
 (5.1) adding the pigment fraction obtained from step (4) to a Sephadex LH-20 column;   (5.2) adding a washing solution to the Sephadex LH-20 column;   (5.3) collecting 3˜5 fractions flowing out from the Sephadex LH-20 column;   (5.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (5.5) mixing the fractions collected in the previous step to form the yellow pigment fraction.   
     
     
         37 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 36 , wherein the washing solution in step (5.2) is Methanol:Acetonitrile=9:1. 
     
     
         38 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the step (6) further comprises the steps of:
 (6.1) adding the yellow pigment fraction obtained from step (5) to a silica gel column;   (6.2) adding a plurality of washing solutions to the silica gel column sequentially;   (6.3) collecting 3˜5 fractions flowing out from the silica gel column;   (6.4) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin; and   (6.5) mixing the fractions collected in the previous step to form the fraction containing monascin and ankaflavin.   
     
     
         39 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 38 , wherein the plurality of washing solutions in step (6.2) are Dichloromethane:Ethanol=95:5, Dichloromethane:Ethanol=9:1, and Dichloromethane:Ethanol=4:1 sequentially. 
     
     
         40 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the step (7) further comprises the steps of:
 (7.1) washing and separating the fraction containing monascin and ankaflavin obtained from step (6) by a C 18  column in combination with a HPLC and a washing solution;   (7.2) collecting 2 fractions flowing out from the C 18  column; and   (7.3) analyzing the fractions collected in the previous step by a HPLC-PDA, and collecting fractions containing monascin and ankaflavin respectively.   
     
     
         41 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 40 , wherein the washing solution in step (7.1) is Methanol:water=85:15. 
     
     
         42 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the specific ratio in step (8) is in a range from 1:1 to 10:1. 
     
     
         43 . The method for manufacturing the composition for lowering blood lipid and elevating high-density lipoprotein according to  claim 31 , wherein the optimum value of the specific ratio in step (8) is 3.56:1.